烟草氧化胁迫相关基因NtOSA1的克隆表达及亚细胞定位分析

发布时间：2018-01-21 03:56

本文关键词： 烟草氧化胁迫相关Abc基因基因克隆表达特性亚细胞定位　出处：《西北植物学报》2016年08期 　论文类型：期刊论文

【摘要】：植物类蛋白激酶Abc1家族(activity of bc1complex)在植物生长发育及响应非生物胁迫中起着重要的作用。该研究通过将拟南芥AtOSA1蛋白氨基酸序列与烟草转录组数据进行比对,利用巢式PCR技术克隆得到1个烟草氧化胁迫相关Abc1家族基因NtOSA1,NtOSA1与拟南芥AtOSA1基因具有72.89%的一致性。NtOSA1基因开放阅读框长度为2 283bp,编码760个氨基酸,含有典型的ABC1结构域、一个激酶结构域、叶绿体定位信号肽和两个跨膜结构域。采用实时荧光定量PCR技术,对NtOSA1基因在烟草不同组织以及氧化胁迫、盐胁迫等处理下的表达分析表明:NtOSA1基因的表达具有组织特异性,主要在叶片中表达;NtOSA1基因在H2O2和NaCl处理后表达量上升,均在处理后6h达到最大值,分别为处理前的1.95和2.69倍。亚细胞定位结果表明,NtOSA1蛋白定位在细胞叶绿体上,与预测结果一致。研究表明,NtOSA1基因参与了烟草抗氧化胁迫和盐胁迫的响应。
[Abstract]:Plant protein kinase Abc1 family activity of bc1 complex. It plays an important role in plant growth and response to abiotic stress. In this study, the amino acid sequences of Arabidopsis AtOSA1 protein were compared with tobacco transcriptional data. A Abc1 gene NtOSA1 of tobacco oxidative stress was cloned by nested PCR technique. The length of NtOSA1 and Arabidopsis AtOSA1 gene was 72.89%. The length of open reading frame of NtOSA1 gene was 2283bpand encoding 760 amino acids. It contains a typical ABC1 domain, a kinase domain, chloroplast localization signal peptide and two transmembrane domains. Real-time quantitative PCR technique is used. The analysis of the expression of NtOSA1 gene in different tissues, oxidative stress, salt stress and other treatments showed that the expression of the NtOSA1 gene was tissue-specific, mainly in the leaves. The expression of NtOSA1 gene increased after treatment with H2O2 and NaCl, and reached the maximum at 6 h after treatment, which was 1.95 and 2.69 times higher than that before treatment, respectively. The NtOSA1 protein was located in the chloroplast of the cells, which was consistent with the predicted results. It was found that the NtOSA1 gene was involved in the responses of tobacco to antioxidant and salt stress.
【作者单位】：西南科技大学生命科学与工程学院;西南科技大学核废物与环境安全国防重点实验室;
【基金】：国家自然科学基金(31500205) 西南科技大学博士研究基金(14zx7153);西南科技大学校长机动费特殊资助(14zx1104) 四川省生物质资源利用与改性工程中心项目(13zxsk02)
【分类号】：Q943.2
【正文快照】： 植物在生长发育过程中不可避免会遭遇到各种逆境胁迫,如高温、低温、干旱、重金属和盐碱等。这些逆境胁迫会使植物氧化还原代谢失衡,导致体内积累大量的活性氧,从而产生氧化胁迫。过量的活性氧能与脂质、蛋白质、DNA以及其他生物大分子发生反应,从而造成机体损伤甚至死亡[1]。

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